Arginine-based PNA microarrays for APOE genotyping

被引：17

作者：

Calabretta, Alessandro ^{[1
]}

Tedeschi, Tullia ^{[1
]}

Di Cola, Gabriella ^{[2
]}

Corradini, Roberto ^{[1
]}

Sforza, Stefano ^{[1
]}

Marchelli, Rosangela ^{[1
]}

机构：

[1] Univ Parma, Dept Organ & Ind Chem, I-43100 Parma, Italy

[2] Biotech Srl, I-43100 Parma, Italy

来源：

MOLECULAR BIOSYSTEMS | 2009年 / 5卷 / 11期

关键词：

PEPTIDE NUCLEIC-ACIDS; STEREOGENIC CENTERS; ALZHEIMER-DISEASE; DNA; HYBRIDIZATION; RECOGNITION; SENSITIVITY; HANDEDNESS; BIOSENSORS; ARRAYS;

D O I：

10.1039/b909912n

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Four modified PNAs containing one chiral monomer bearing two arginine-derived side chains, with the correct configuration for specific and stable DNA binding, were synthesized, complementary to two DNA tracts in the APOE gene containing SNPs related to the insurgence of Alzheimer's disease. PNA binding performances were first tested in solution against complementary and mismatched oligonucleotides by measuring melting temperatures, and showed high specificity in SNP recognition. In order to set up a new diagnostic platform for APOE genotyping, PNA microarrays were then developed with the synthesized modified PNAs. PNA probe deposition protocols on microarrays were optimized in order to minimize cross-contamination due to carry over. The microarrays obtained by arginine-based PNA deposition were incubated with complementary and mismatched oligonucleotides, showing excellent mismatch recognition on the microarray platform. The specificity of the microarrays was finally tested with oligonucleotide mixtures simulating the real genotype profiles. Six different hybridisation patterns related to six different genotypes in the APOE gene were found to be clearly distinct in microarray experiments, demonstrating the potential of this approach for highly specific genetic analysis.

引用

页码：1323 / 1330

页数：8

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